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Guillermo Ameer
Northwestern University
Corresponding Author
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Efficient manipulation of cell fate is important for regenerative engineering applications. Lineage-specific differentiation of stem cells is particularly challenging due to their inherent plasticity. Engineered topographies may alter cellular plasticity through contact guidance. However, the ability to rationally design topographies to regulate phenotypic outcomes has been hindered in part by the lack of tools to quantify nanoscale chromatin structure reorganization in live cells. Herein we use micropillars, molecular, and nanostructural quantification tools to investigate how nuclear morphology in human mesenchymal stem cells (hMSCs) affects chromatin conformation and osteogenic differentiation. We show that micropillar-induced contact guidance is transduced via the cytoskeleton and impacts nuclear architecture, lamin A/C multimerization, histone modifications, and the 3-D conformation of chromatin within packing domains, a key regulator of transcriptional responsiveness. Micropillars repressed expression of genes associated with developmental processes and enhanced lineage-specific responsiveness, thereby decreasing cell plasticity and off-target differentiation, and facilitating osteogenic differentiation of hMSCs. Altogether, these findings reveal that chromatin reprogramming through contact guidance-induced nuclear deformation can be an efficient way to manipulate cell fate.
Keywords
stem cells, biotechnology, genetics
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There is NO Competing Interest.
This is a list of supplementary files associated with this preprint. Click to download.
- TableS1MicropillarvsFlatStemMSCsDifferentialExpressionSignificant.xlsx
Supplementary Table S1
- TableS2GeneOntologyMetascapePillarvsFlatDowregulatedGenesStemXInducedConditions.xlsx
Supplementary Table S2
- TableS3Top20OntologyGeneSetsFlatXMicropillars.csv
Supplementary Table S3
- TableS4GeneOntologyMetascapeInducedFlatvsStemFlatbyPandLFC.xlsx
Supplementary Table S4
- GuillermoA.Ameer0209AmeerTableS5Rawdataforplottingfiguresinmain.xlsx
Supplementary Table S5
- Nat.BMEMicropillarsSI.docx
Chromatin Reprogramming via Contact Guidance-Induced Nuclear Deformation Promotes Stem Cell Differentiation
- AmeerRS.pdf
Reporting summary flat
- GuillermoA.Ameer0209AmeerSupplementaryInformation.docx
SI
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A new preprint design is coming!
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This is a preprint, a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Article
Guillermo Ameer
Northwestern University
Corresponding Author
Badges
Prescreen
This manuscript passed our Prescreen assessment
Complete author information
Appropriate declaration statements
Potential risks to human health
Prescreen
History
CURRENT STATUS: Posted
Version 1
Posted 04 Feb, 2021
No community comments so far
Metrics
Comments: 0
PDF Downloads: ...
HTML Views: ...
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Efficient manipulation of cell fate is important for regenerative engineering applications. Lineage-specific differentiation of stem cells is particularly challenging due to their inherent plasticity. Engineered topographies may alter cellular plasticity through contact guidance. However, the ability to rationally design topographies to regulate phenotypic outcomes has been hindered in part by the lack of tools to quantify nanoscale chromatin structure reorganization in live cells. Herein we use micropillars, molecular, and nanostructural quantification tools to investigate how nuclear morphology in human mesenchymal stem cells (hMSCs) affects chromatin conformation and osteogenic differentiation. We show that micropillar-induced contact guidance is transduced via the cytoskeleton and impacts nuclear architecture, lamin A/C multimerization, histone modifications, and the 3-D conformation of chromatin within packing domains, a key regulator of transcriptional responsiveness. Micropillars repressed expression of genes associated with developmental processes and enhanced lineage-specific responsiveness, thereby decreasing cell plasticity and off-target differentiation, and facilitating osteogenic differentiation of hMSCs. Altogether, these findings reveal that chromatin reprogramming through contact guidance-induced nuclear deformation can be an efficient way to manipulate cell fate.
Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6
There is NO Competing Interest.
This is a list of supplementary files associated with this preprint. Click to download.
- TableS1MicropillarvsFlatStemMSCsDifferentialExpressionSignificant.xlsx
Supplementary Table S1
- TableS2GeneOntologyMetascapePillarvsFlatDowregulatedGenesStemXInducedConditions.xlsx
Supplementary Table S2
- TableS3Top20OntologyGeneSetsFlatXMicropillars.csv
Supplementary Table S3
- TableS4GeneOntologyMetascapeInducedFlatvsStemFlatbyPandLFC.xlsx
Supplementary Table S4
- GuillermoA.Ameer0209AmeerTableS5Rawdataforplottingfiguresinmain.xlsx
Supplementary Table S5
- Nat.BMEMicropillarsSI.docx
Chromatin Reprogramming via Contact Guidance-Induced Nuclear Deformation Promotes Stem Cell Differentiation
- AmeerRS.pdf
Reporting summary flat
- GuillermoA.Ameer0209AmeerSupplementaryInformation.docx
SI
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